农业科学与技术:英文版
農業科學與技術:英文版
농업과학여기술:영문판
Agricultural Science & Technology
2012年
1期
177-180
,共4页
刘晶晶%胡美英%任珍珍%李亚楠
劉晶晶%鬍美英%任珍珍%李亞楠
류정정%호미영%임진진%리아남
小菜蛾%化学感受蛋白1%原核表达
小菜蛾%化學感受蛋白1%原覈錶達
소채아%화학감수단백1%원핵표체
P. xylostella%PlxyCSP1%Prokaryotic expression system
[目的]研究半胱氨酸(cys58)对小菜蛾(爿utellaxylosteff由化学感受蛋白I(P1xyCSP1)与农药化合物结合特性的影响。[方法]利用重叠延伸PCR法,将PlxyCSP1中Cys58突变成色氨酸(Trp58),获得突变体P1xyCSPI—M2。构建表达载体表达突变蛋白,并进行WesternBlot检测。[结果]构建了突变基因的原核蛋白表达载体pET32a—P1xyCSP1-M2,并表达35kDa融合蛋白【结论]利用重叠延伸PCR法成功在原核系统中表达了PlxyC—SP1突变蛋白。
[目的]研究半胱氨痠(cys58)對小菜蛾(爿utellaxylosteff由化學感受蛋白I(P1xyCSP1)與農藥化閤物結閤特性的影響。[方法]利用重疊延伸PCR法,將PlxyCSP1中Cys58突變成色氨痠(Trp58),穫得突變體P1xyCSPI—M2。構建錶達載體錶達突變蛋白,併進行WesternBlot檢測。[結果]構建瞭突變基因的原覈蛋白錶達載體pET32a—P1xyCSP1-M2,併錶達35kDa融閤蛋白【結論]利用重疊延伸PCR法成功在原覈繫統中錶達瞭PlxyC—SP1突變蛋白。
[목적]연구반광안산(cys58)대소채아(장utellaxylosteff유화학감수단백I(P1xyCSP1)여농약화합물결합특성적영향。[방법]이용중첩연신PCR법,장PlxyCSP1중Cys58돌변성색안산(Trp58),획득돌변체P1xyCSPI—M2。구건표체재체표체돌변단백,병진행WesternBlot검측。[결과]구건료돌변기인적원핵단백표체재체pET32a—P1xyCSP1-M2,병표체35kDa융합단백【결론]이용중첩연신PCR법성공재원핵계통중표체료PlxyC—SP1돌변단백。
[Objective] This study aimed to explore the effects of cysteine (Cys58) on the characteristics of PlxyCSP1 in P. Iostella combined with pesticide compounds. [Method] Cys58 of PlxyCSP1 was mutated into Trp58 by using overlap extension PCR method, and PlxyCSP1-M2 mutant was obtained. Expression vector was con- structed and the protein was detected by western blot. [Result] Expression vector pET32a-PlxyCSP1-M2 was constructed to express the 35kDa weight protein. [Conclusion] PlxyCSP1 mutant protein has been expressed successfully in prokaryotic ex- pression system.